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. 2011 Mar 14;208(3):577–592. doi: 10.1084/jem.20100619

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© 2011 Wang et al.

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Figure 5. — Immobilized VISTA-Ig fusion protein inhibited CD4⁺ and CD8⁺ T cell proliferation. CFSE-labeled CD4⁺ and CD8⁺ T cells were stimulated by plate-bound α-CD3 together with co-absorbed VISTA-Ig or control-Ig protein at the indicated ratios. (A) Representative CFSE dilution profiles. (B) The percentage of CFSE-low cells was quantified and shown as mean ± SEM. (C) As in A, but using CD4⁺ T cells from PD-1–deficient mice. (D) Wild-type CD4⁺ T cells were activated in the presence of VISTA-Ig or control-Ig for 72 (i) or 24 h (ii–iv). 24-h preactivated cells were harvested and restimulated under specified conditions for another 48 h. (ii) Preactivation with VISTA-Ig and restimulation with α-CD3. (iii) Preactivation with α-CD3 and restimulation with control-Ig or VISTA-Ig. (iv) Preactivation with control-Ig or VISTA-Ig and restimulation with the same Ig protein. Cell proliferation was analyzed at the end of the 72-h culture. Duplicated wells were analyzed for all conditions. Shown are representative results from at least four experiments.